RESUMO
BACKGROUND: Cell-free DNA (cfDNA) analysis using peripheral blood represents an exciting, minimally invasive technology for cancer diagnosis and monitoring. The reliability of testing is dependent on the accuracy and sensitivity of specific molecular analyses to detect tumor-associated genomic variants and on the quantity and quality of cfDNA available for testing. Specific guidelines for standardization and design of appropriate quality programs focused specifically on cfDNA isolation are lacking, as are standardized quality control reagents. CONTENT: This report describes and illustrates quality control and quality assurance processes, supported by generation of in-house quality control material, to ensure the reliability of the preanalytical phase of cfDNA analysis. SUMMARY: We have developed a robust quality program to support high-volume automated cfDNA extraction from peripheral blood by implementing processes and procedures designed to monitor the adequacy of specimen collection, specimen stability, efficiency of cfDNA extraction, and cfDNA quality.
Assuntos
Coleta de Amostras Sanguíneas/normas , DNA Tumoral Circulante/isolamento & purificação , Serviços de Laboratório Clínico/normas , Guias como Assunto , Neoplasias/diagnóstico , DNA Tumoral Circulante/genética , Serviços de Laboratório Clínico/organização & administração , Análise Mutacional de DNA , Humanos , Mutação , Neoplasias/sangue , Neoplasias/genética , Controle de Qualidade , Melhoria de Qualidade , Reprodutibilidade dos TestesRESUMO
Digital microfluidics (DMF), a technique for manipulation of droplets, is a promising alternative for the development of "lab-on-a-chip" platforms. Often, droplet motion relies on the wetting of a surface, directly associated with the application of an electric field; surface interactions, however, make motion dependent on droplet contents, limiting the breadth of applications of the technique. Some alternatives have been presented to minimize this dependence. However, they rely on the addition of extra chemical species to the droplet or its surroundings, which could potentially interact with droplet moieties. Addressing this challenge, our group recently developed Field-DW devices to allow the transport of cells and proteins in DMF, without extra additives. Here, the protocol for device fabrication and operation is provided, including the electronic interface for motion control. We also continue the studies with the devices, showing that multicellular, relatively large, model organisms can also be transported, arguably unaffected by the electric fields required for device operation.